Validated using a knockout cell line

Anti-Nestin antibody [2C1.3A11] (ab18102)

Reviews (1)Q&A (1)References (4)

Overview

  • Product name
    Anti-Nestin antibody [2C1.3A11]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [2C1.3A11] to Nestin
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Rat
  • Immunogen

    A 150 amino acid fragment from the cloned human Nestin.

  • General notes


    This highly specific antibody to human nestin can aid in characterizing progenitor cells differentiating into distinct lineages, in enhancing the therapeutic potential of human neural stem/progenitor cells in the treatment of CNS diseases or injury, and in identifying neuroepithelial tumor cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab18102 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/5000. Predicted molecular weight: 177 kDa.
Flow Cyt 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function
    Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells.
  • Tissue specificity
    CNS stem cells.
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Developmental stage
    Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
  • Post-translational
    modifications
    Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
  • Information by UniProt
  • Database links
    • Alternative names
      • ESTM 46 antibody
      • FLJ 21841 antibody
      • FLJ21841 antibody
      • Intermediate filament protein antibody
      • Nbla00170 antibody
      • nes antibody
      • NEST_HUMAN antibody
      • Nestin antibody
      see all

    Images

    • Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      All lanes : Anti-Nestin antibody [2C1.3A11] (ab18102) at 1/5000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : NES knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 177 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab18102 observed at 300 kDa. Red - loading control, ab176560, observed at 50 kDa.

      ab18102 was shown to recognize NES in wild-type HAP1 cells as signal was lost at the expected MW in NES knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab18102 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunofluorescence staining of U251 cells (glioblastoma cell line) stained using ab18102(green) and bisbenzimide (blue)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of migrating tumor cells labelling Nestin with ab18102 at 2µg/ml. Cytoplasm and membrane positive staining shown.

    • Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      All lanes : Anti-Nestin antibody [2C1.3A11] (ab18102) at 1/1000 dilution

      Lane 1 : Total rat brain protein at 5 µg
      Lane 2 : Human CNS progenitor cell protein at 1 µg
      Lane 3 : Human CNS progenitor cell protein at 5 µg

      Predicted band size: 177 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of xenograft tumor cells labelling Nestin with ab18102.

    • Flow Cytometry - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Flow Cytometry - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Overlay histogram showing SH-SY5Y cells stained with ab18102 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18102, 1/200 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry of Anti-Nestin on formalin fixed paraffin embedded prostate nerve tissue using ab18102

    References

    This product has been referenced in:
    • Ferrer CM  et al. An inactivating mutation in the histone deacetylase SIRT6 causes human perinatal lethality. Genes Dev 32:373-388 (2018). Read more (PubMed: 29555651) »
    • Jaraíz-Rodríguez M  et al. A Short Region of Connexin43 Reduces Human Glioma Stem Cell Migration, Invasion, and Survival through Src, PTEN, and FAK. Stem Cell Reports 9:451-463 (2017). Read more (PubMed: 28712848) »
    See all 4 Publications for this product

    Publishing research using ab18102? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Flow Cytometry abreview for Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Neural Progenitor cells (from human ES cells))
    Specification
    Neural Progenitor cells (from human ES cells)
    Preparation
    Cell harvesting/tissue preparation method: Cell Dissociation Buffer
    Sample buffer: FACS buffer (0.2%BSA, 0.1%sodium azide in PBS)
    Fixation
    Paraformaldehyde
    Permeabilization
    Yes - 70% Methanol (ice cold)
    Gating Strategy
    FSC vs SSC on negative control sample
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 03 2012

    Question

    Yes, that would be for the nestin antibodies only. (ab6320 & ab18102) If you can find any specific FACS protocol for one of these, then it may be decisive in my choice. Up to know, I do not have any information for choosing between them. Thus I am waiting for your answer. Anyway, if you cannot find any protocols,then I will choose one of them by chance. Thanks in advance, I hope this helps

    Read More
    Answer

    I am please to be able to share the following protocol for ab18102for Flow cytometry: Fix cells with 4% Paraformaldehyde for 30 min at room temperature (RT) Wash twice with 1% PBS at 1500rpm/5min/RT Permeabilize cells with 0.02% Triton X-100 in 1%PBS + 1% BSA for 15 min/RT Then add rabbit nestin ab (1:400 dilution) for 1hr/RT Wash twice Add specific secondary ab for 1hr/RT Wash twice Flow reading I hope this information will be helpful. Please do not hesitate to contact me if I can be of further assistance,

    Read More

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